Steroid compounds and methods for their manufacture



2,814,632 1C p ted Nov. 26, 1957 2.8 am s runom COMPQUNDS. METHODS FOR 'runm MANUFACTURE Alexander L. Iflussbaum, Bloomfield, N. J., assignor t Sclie'riug Corporation, "Bloomfield, N. 1., acorporation ofNew-Jersey rid Draw-tee Ap lica io Febru ry 2,1 @257.

are]; on 487 ZIClaims. (Cl. 26071-39145) wherein. X. s a e b of t e gro p onsist ng of hyrones. and fluor n Y is a member of the gro p n-- stine of satlt s a. membe of e g o c s g Q a d. aey radicals of lower alkanoic acids.

re. tqd h he comp u ds of e en rmula 'are adrenocorticoid in their action are. thus useful in the treatment or inflammation, arthritic diseases and certain other collagen diseases such as disseminated lupus, polyarteri tis and the like. The thiowcompounds of my invention as, compared; with their oxygen analogues, exhibit. a greater diuretic action. Thus, following administration of. thesethio-compounds, there is relativey ittle So u ent o a d s m t d dem w i isjso common with many of the,adrenocorticalhormones and, corticoids heretofore. in. use. My new compounds are thus preferentially employed. especially when adrenacorticoid deficiencies are accompanied. by cardiac insufliciencies, The new compounds of. this invention byexerting agrnarked, diuresis are useful adjuncts inthetreatment of 'congestiye heart failure.

Inaddition, the free thiol-compounds of my, invention form insoluble precipitates with heavy metals such as leed and. may be used in the qualitative or quantitative es imatio uch h a y et s,

The compounds of my invention may readily be pre pared: from the known compounds, prednisone and prednisolone. The Zl-h-yd-roxy group of these compounds is e sterified with the acid chloride of any alkylor aryl;sulfonic acid such as methyl-, ethyl-, benzene-, or p toluenesulfonic acid. The reaction may be carried out irlr the presence of a base such as pyridine or triethylamine, and with or without a solventsuch.as chloroform,

dime y tormamid i xane or the l ke. The re l n ester is converted to the 21 iodide Qr Zl-bromideby means o s dium iodi o odium romide in he known mann r. The 21-h 1i o ob a d i of ufficient p rity to be used in subsequent transformations without further purification. The 2l-iodide of prednisone or prednisolone and a salt, preferably alkali metal, of a ,thiol acid such as thiolacetic or thiolpropionic are heated together in a preferably refluxing solvent such as acetone or methanol to yield the new compounds 1,4-pregnadiene- 17ot-Ol-21-thl0l-3,11,20-lII'l0l16 21-acetate (or .propionate) when the starting material is prednisone; and 1,4- P esn e 7e.- iol. 2 -thio1 3,2.0 dione. 21 cet te (o pr n onat r m prednisoloue, 1 pre n. i ne-Hit-01 21-t io -3.l.l, .0-trio Z -a etate (or pr p Qnate) may o be Prep r fro th corr spondin hydroxy compound by oxidation with agents such as chromium d oxid (w th. or without py n present), Sodi m di romate, N rom a mide, N- romosuce imid a d the like,

T f ee 2,14H compounds of y invention may e prepared from the thiolesters described above by sub.- j ecting said esters to the action of Flavobacterium, dehydrogenwns var. hydrolyticum as analogously described in the "co-pending application of William Channel, Serial No. 458,661, filed September 27, 1954. Thus, 1,4- pregnadiene l7e ol-21-.thiol 3,l1,20 trione and 1,4,=pregnadiene-l1,8,17a-dio1-21-thi0l 3,20 dione are. obtained from their corresponding 21-thiolesters.

To prepare the compounds of my invention in which there is a 9a -fluoro moiety, similar methods to the above are applicable starting with either 9u-fluoroprednisone or 9o c-fluoroprednisolone.

The following examples set forth methods for preparing the compounds embraced by my invention; however, it is to be understood that the examples are presented for illustrative purposes and my invention is limited only as set forth. in the appended claims.

EXAMPLE, 1

1,4-pregnadiene-1 1 13,1 7oa-dz'0l-21--thi0l-3,20-dione 21- acetate A. 1,4-PREGNADIENE-l1fl,17a,21-TRIOL-3,20-DIONE 2l-P-TOLUE NESULFONATE To a solution of 10 grams of prednisolone in 57.5 ml. of pyridine, frozen in a Dry Ice-acetone bath, is added, a chilledsolution of 5.9 grams of .p-toluenesulfonyl chloride in 47 ml. of methylene chloride. The mixture is maintained in a Dry Ice-acetone bath for 2 hours and then allowed to stand at 18 C. for 20 hours. Cold ether (500.ml.) is added and the resulting solution is extracted in turn with water, dilute hydrochloric acid, dilute sodibicarbonate, and finally water. The non-aqueous solution is dried over sodium sulfate, filtered, and concentrated to dryness in vacuo. The residue is crystallized from methanol, yielding 13.4 grams. of l,4-pregnadiene-. 1lfl,1766,21-t1'l0lr3,20-dl0116 21-p-toluenesulfonate, M. P. l28-135 C.; ultraviolet absorption at 228- my. (e=21 ,,300), shoulder at 242 mu (e=15,000).

B. 21-IODO-1,4-PREGNADIENE-11B,17a-DIOL-3,20-DIONE 10 grams of the 21-p-toluenesulfonate of prednisoloneprepared above is dissolvedin 50 ml, of acetone, and 10 grams of sodium iodide in 70 ml; of acetone is added. The mixture is refluxed for seven minutes duringv which time aheavy precipitate forms, Themixtureis concentratedto dryness in vacuo and 300 ml. of 0.33 N'sodiurn thiosulfate-is. added to the residue. 2,1-iodo- 1,4-pregnadiene-l1,8,17ot-diol-3,20-dione which precipitatesis filtered. and dried to, yield 8.2,grams, M. P. -170 C; dec.

C. 1,4PREGNADIENE-11B,17n-DIOL-21-THIOL-3,20- DIONE 21-ACETATE A solution of grams of 21-iodo-prcdnisolone (from 1B) in 250 ml. of acetone is added to a suspension of 1.7 grams of potassium tln'olacetate in 200 ml. of acetone. The mixture is refluxed for 2 hours, and concentrated in vacuo almost to dryness. One liter of water is added, and the resulting suspension is filtered and recrystallized from methanol to yield 3.5 grams of l,4-pregnadiene- 11,8,l7a-di0l-2l-thiol-3,20-dione 21-acetate, M. P. 237- 246 C. [a] =+126.7 (CHClg).

EXAMPLE 2 1,4 pregnadiene 17a ol 21 thiol 3,11,20 trione 21-acetate 500 milligrams of 1,4-pregnadiene 115,170: diol-2lthiol-3,20-dione ZI-acetate is dissolved in 50 ml. of pyridine and added to a mixture consisting of 310 mg. of chromium trioxide in 35 ml. of pyridine. The reaction mixture, after standing overnight at room temperature, is poured into dilute hydrochloric acid and extracted with methylene chloride. The organic layer is extracted with dilute sodium bicarbonate and then a suflicient number of times with water until the aqueous extracts are neutral to litmus. The non-aqueous solution is dried over sodium sulfate, filtered, and concentrated to dryness in vacuo. The residue is crystallized from ether to yield 376 mg. of l,4-pregnadiene-l7a-ol-21-thiol-3,l1,20-trione ZI-acetate.

Alternatively, the compound of this example may be prepared as follows:

A. 1,-1-PREGNriDIENE-l7mZ1-DIOL-3.ILZO-TRIONE 21-P-TOLUENESULFONATE A solution of 10 grams of prednisone in 57 ml. of pyridine is reacted with p-toluenesulfonate in the manner described in Example 1A, yielding 1,4-pregnadiene-17e,2ldiol-3,ll,20-trione ZI-p-toluenesultonate, which is crystallized from methanol.

B. 2l-IODO-1,4-PREGNADIENE-1Tot-OL-S,11,20TRIONE 10 grams of the 21-p-toluenesulfonate of prednisone prepared above is reacted with sodium iodide in acetone as described in Example 1B, yielding 21-iodo-1,4-pregnadiene-17a-ol-3,11,20-trione.

c. l,4-PREGNADIENE-l7a-OL-2]-THIOL-3,11,20-TRIONE 21-ACETATE A solution of 5 grams of 2l-iodo-prednisone prepared in Procedure B of this example, is reacted with potassium thiolacetate in the manner described in Example 1C. The resulting product, l,4-pregnadiene-]7e-ol-21-thiol- 3,1],20-trione ZI-acetate is recrystallized from ether.

EXAMPLE 3 1,4-pregnadielze-115,17oc-di0l-2I-thi0l-3,20-di0ne An inoculum is prepared as follows:

A culture of Flavobacteritmz dehydrogenans var. hydrolyticum (which organism is on deposit at Rutgers University under the designation Flavobacterium dehydrogenans 130) is grown for 24 hours with forced aeration at 2830 C. in a medium consisting of a tap water solution containing 1% of condensed fish solubles (50% solids), 0.1% of Basamin yeast hydrolysate, 0.8% of disodium phosphate heptahydrate. 0.4% of potassium dihydrogen phosphate and 0.05% of a defoamer which has been sterilized at 125 C. under pressure for 2% hours.

Similarly 500 ml. of the above medium is prepared, sterilized and distributed equally among five, 300 ml. Erlenmeyer flasks. To each flask is added 2 ml. of the inoculum described above and the flasks and contents are shaken on a shake table for 16 hours at 2830 C. To each flask is then added 200 mg. of 1,4-pregnadienellfl,l7e-dicl-21-thiol-3,20-dione Zl-acetate in 2 ml. of acetone, and the fermentation is continued. After 24 hours, a further addition of 200 mg. of steroid is added to each flask, and shaking is continued another 24 hours. The combined brei are then extracted twice with chloroform. The chloroform solution is treated with activated charcoal, filtered, and evaporated. The residue is crystallized from acetone-hexane to give 1,4-pregnadiene- 11,8, l7oc-dlOl-2 l-thio1-3,20-dione.

EXAMPLE 4 1,4-pregnadiene-1 7ot-0l-21-lhi0l-3J1,20-tri0ne A solution of 1,4-pregnadiene-17a-ol-21-thi0l-3,11,20- trione 21-acetate in 2 ml.,of acetone is subjected to the action of a culture of Flavobacterium dc/zydrogenans var. hydrolytz'cum in the manner described in Example 3 to yield l,4-pregnadiene-l7a-ol-2l-thiol 3,11,20 trione, which is crystallized from acetone-hexane.

EXAMPLE 5 9m fluoro 1,4 pregnadiene 1. 3,170: dial 21 thiol- 3,20-di0ne ZZ-acetate A. 2l-IODO-OdFLUORO-1,4-PREGNADIENE-11B,17a-DIOL- 3,20-DIONE 1.6 grams of 9a-fluoroprednisolone ZI-methanesulfonate (M. P. 22l-224 C. doc.) is dissolved in 68 ml. of acetone and to this is added a solution of 5.52 g. of sodium iodide in acetone. The resulting solution is refluxed for 15 minutes, during which time a heavy precipitate forms. This suspension is filtered while hot and the filtrate containing 21-iodo 9a fluoro-1,4-pregnadiene- 11B,l7a-diol-3,20-dione is used directly in the next step.

B. 9a-FLUORO-IAPREGNADIENE-I1B.17r1-DIOL-21- THIOL'BZO-DIONE 21-ACETATE The hot acetone solution of 21-iodo-9u-fiuoro-L4- pregnadiene-llB,l'7a-diol-3,20-dione is poured immediately into a suspension of 6.29 g. of potassium thiolacetate in ml. of acetone. The mixture is refluxed for 2 hours, and then concentrated in vacuo. Water (350 ml.) is added to the residue and the precipitate is filtered and dried yielding 1.43 g. of 9a-fluoro-1,4-pregnadiene- 115,17a-di0l-2l-thiol-3,20-dione 2l-acetate, M. P. 225 233 C.

EXAMPLE 6 9a-fluor0 1,4 pregnadiene 17a ol 21 thiol 3,11,20-

trione 21 -acetale 500 milligrams of the product of Example 5 is dissolved in 50 ml. of pyridine and oxidized with chromium trioxide in the manner described in Example 2, to yield 9a-fluoro-l,4-pregnadiene-17a-0l-2l-thi0l-3,11,20- trione 21-acetate.

EXAMPLE 7 9a-flu0r0 1,4 pregnadfene 17a ol 21 thiol 3,11,20- trione 21 -pr0pi0nate A. 9aFLUORO-1.4-PREGNADIENE-17a.21-DIOL-3,11,20- TRIONE 21-METHANESULFONATE 2.5 grams of 9oc-fll1010 1,4 pregnadiene-l7tx,2l-diol- 3,11,20-trione is dissolved in 75 ml. of methylene chloride and cooled to 0 C. To the cooled solution is added 5 m1. of pyridine and 10 ml. of methanesulfonyl chloride. The reaction mixture is held at 0 C. for 2 hours, and then at room temperature for 20 hours. Ethyl acetate (250 ml.) is added and the organic solution is Washed in turn with dilute hydrochloric acid, dilute aqueous sodium carbonate, and finally water. The ethyl acetate solution is dried over sodium sulfate, filtered, and concentrated in vacuo to incipient crystallization. Upon cooling and filtration there is obtained 1.7 g. of 9e-flUoro-L4-pregnadiene-l 7a,2l-di0l-3,l 1,20-dione ZI-methanesulfonate.

B. 2]-IODO-9ct-FLUORO-1.4-PREGNADIEND17o-21-DIOL- 3,11,20-TRIONE 1.5 grams of 9u-fluoroprednisone ZI-rnethanesulfonate in 65 ml. of acetone is reacted with 5.45 grams of sodium iodide in the manner described in Example 5A. The hot filtrate containing 2l-iodo c fiuoroprednisone is used directly in the next step.

5 C. 9a-FLUORO-l,4-PREGNADIENE-17a-OL21-THIOL- 3,11,20-TRIONE 21-PROPIONA'1E The hot solution of 21-iodo-9a-fluoro-1,4-pregnadiene- 17oc,21-di0l-3,11,20-11101'16 in acetone is added directly to a suspension of 6.7 g. of potassium thiolpropionate in 75 ml. of acetone and refluxed for 2 hours. The mixture is concentrated in vacuo, and 350 ml. of water is added to the residue. The precipitate is filtered and dried, yielding 9a-fluoro-1,4-pregnadiene-17a-ol-2l-thiol- 3,11,20-trione 21-propi0nate.

EXAMPLE 8 9a-fluoro-1,4-pregnadiene-11,3,17u-di0l-21-thiol- 3,20-dine 200 milligrams of 90L-fillOI'O-1lfi,170L-di01-21-thlOl-3,20- dione 2l-acetate (product of Example 5) is subjected to the action of a culture of Flavobacterium dehydrogenans var. hydrolyticum in the manner described in Example 3, to yield 9a-fluor0-1,4-pregnadiene-11,3,17a-diol-21-thiol-3, ZO-dione which is crystallized from acetone-hexane.

200 milligrams of 9a-fluoro-17a-ol-21-thiol-3,11,20-trione 21-propionate (product of Example 7) is subjected to the action of a culture of Flavobacterium dehydrogenans var. hydrolyticum in the manner described in Example 3 to yield 9a-fluoro-1,4-pregnadiene-17a-ol-21-thiol-3,11,20- trione.

I claim:

1. Compounds having the structural formula CHRSR wherein X is a member of the group consisting of hydrogen and fluorine; Y is a member of the group consisting of O and (H, OH) and R is a member of the group consisting of H and acyl radicals of lower alkanoic acids.

2. A 21-l0wer alkanoate of 1,4-pregnadiene-11 5,1701- diol-2l-thiol-3,20-dione.

3. A 21-lower alkanoate of 1,4-pregnadiene-17a-ol-21- thiol-3,11,20-trione.

4. A 21-lower alkanoate of 9u-fluoro-1,4-pregnadiene- 1 1p,17a-diol-21-thiol-3,20-dione.

5. 1,4-pregnadiene-115,17ct-di0l-21-thiol-3,20-dione 21- acetate.

6. 1,4 pregnadiene 17cc o1 21 thiol 3,11,20 trione 21-acetate.

7. 9oz fluoro 1,4 pregnadiene 11 3,17 diol 21- thiol-3,20-dione 21-acetate.

8. 9oz fluoro 1,4 pregnadiene 17a o1 21 thiol- 3,11,20-trione 21-propionate.

9. 1,4-pregnadiene-1 1 8,17a-diol-21-thio1-3,20-dione.

10. 1,4-pregnadiene-17a-ol-21-thiol-3,11,20-trione.

11. 9a fluoro 1,4 pregnadiene 115,170 diol 21- thiol-3,20-dione.

12. In the process for preparing steroid compounds of the formula CHzSR Y i X wherein X and Y are as above defined and Z is a halogen radical having an atomic weight greater than chlorine with a salt of a thiol-lower alkanoic acid.

13. The process of claim 12 wherein the reaction is carried out in an inert organic solvent.

14. The process of claim 13 wherein the salt of the thiol-lower alkanoic acid is potassium thiolacetate.

15. The process according to claim 14 wherein the starting steroid is 21-iodo-1,4-pregnadiene-17aaol-3J1,20-trione.

16. The process according to claim 14 wherein the starting steroid is 21-iodo-9u-fluoro-1,4-pregnadiene-17aol-3,1 1 ,20-trione.

17. The process according to claim 14 wherein the starting steroid is 2l-iodo-1.,4-pregnadiene-11fi,17a-di0l-3, 20-dione.

18. The process of claim 17 including the step of 0xidizing the llfi-hydroxyl group to a ketone.

19. The process of claim 14 wherein the starting steroid is 21-i0do-9a-fluoro-1,4-pregnadiene-11p,17a-diol-3,20-dione.

20. The process of claim 19 including the step of oxidizing the llp-hydroxyl group to a ketone.

21. The process of claims 12-14 including the step of hydrolyzing the 21-thiolester to form a 21-thiol.

No references cited. 

1. COMPOUNDS HAVING THE STRUCTUAL FORMULA 